CN101347617B - Refined polyvalent anti-snake poison lyophilized blood serum and using method - Google Patents
Refined polyvalent anti-snake poison lyophilized blood serum and using method Download PDFInfo
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- CN101347617B CN101347617B CN2006100109334A CN200610010933A CN101347617B CN 101347617 B CN101347617 B CN 101347617B CN 2006100109334 A CN2006100109334 A CN 2006100109334A CN 200610010933 A CN200610010933 A CN 200610010933A CN 101347617 B CN101347617 B CN 101347617B
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Abstract
Purified multivalent and lyophilized antivenin and a use method belong to antibody immunity serums that aim at more than two antigens. The invention aims at preparing the multivalent antivenin by a plurality of lyophilized antivenins and the use method. The invention is characterized in that: the antigen-binding fragment F(ab)2 has 60 to 80 percent of lyophilized antiserum, the unit is mg per person, 342.5 to 1035 of vipers, 1177 to 3182 of agkistrodon acutus, 456 to 666 of adders, 1419 to 2128 of king cobras, 220 to 331 of coral snakes, 880 to 1320 of gold banded kraits, 3188 to 4782 of cobras are combined; the optimized combination is as follows: 635 of vipers, 1959 of agkistrodon acutus, 561 of adders, 1774 of king cobras, 276 of coral snakes, 1100 of gold banded kraits and 3985 of cobras. Blood circulation snake venom antiserum and nerves snake venom antiserum are respectively filled and measured. The use method is as follows: the antivenin that is diluted by sodium chloride and an anti-allergic agent, such as chlorphenamine maleate, are injected into muscle or a vein. The antivenin is prepared by only seven antivenins, and can effectively and in time treat injuries caused by various poisonous snakes in our country when the antiserum is directly injected in condition of the undetermined variety of poisonous snakes, and the invention reaches the advanced world level of the antivenin.
Description
Affiliated technical field
The invention belongs to the immune serum that contains antibody, particularly contain immune serum at two or more antigenic antibody.
Background technology
The unit price antivenin of certain snake venom preparation, the strongest to being used to prepare the neutral specificity of the sero-fast snake venom of this kind, take second place to belonging to other snake venom that same section belongs to, and poorer even do not have to not equal snake venom.When using clinically, must make a definite diagnosis the Serpentis kind that is snapped after, the significant curative effect of competence exertion.For under the situation of not determining which kind of venom, can neutralize effectively that same section belongs to or the snake venom of equal genus not with a kind of calmette's serum, the scientist selects multiple snake venom to prepare antigen to prepare multivalence anti-venin serum to animal immune.Multivalence anticrotalus serum for example produced in USA, select the hybrid antigen of US West's Pedicellus et Pericarpium Trapae speckle crotalin (Crotalusatrox), east Pedicellus et Pericarpium Trapae speckle crotalin (Crotalus adamanteus), the terrified subspecies snake venom (Crotalus durissusterrificus) of rattle snake and spearhead agkistrodon halyx pallas venom (Bothrops atrox) for use, use the horse immunity, be prepared from by horse blood slurry, in the multivalence anti-venin serum energy of making by these four kinds of snake venom and 17 kinds of snake venom among the Crotalidae Crotalidae.But, this serum same section poison of being born in the year of snake that still is confined to neutralize, and this serum only bites at this area rattle snake and has curative effect.Though China prepares multiple unit price antivenin,, still there is not polyvalent serum to endangering the snakebite due to the big zonal various poisonous snakes.
Before this, inventor's number of filing an application 200610010908.6 " method of refined polyvalent anti-snake poison lyophilized blood serum " patent application, comprise and use Agkistrodon, Agkistrodon halys, Trimeresurus mucrosquamatus (Cantor)., Trimeresurus stejnegeri, adder, Agkistrodon, Bungarus fasciatus, Naja, Ophiophagus hannan (Cantor) venom as immunizing antigen, with micro-progressive immune animal after membrane filtration degerming dialysis, the detoxification, after crossing column chromatography purification, gastric enzyme digestion-ammonium sulfate precipitation collects antibody, desalination, lyophilizing; And tire with mouse experiment assessment snake venom virulence half mortality LD50 and antivenin.
Summary of the invention
The objective of the invention is to prepare multivalence anti-venin serum, and propose when clinical treatment, to use the method for this serum with the multiple snake poison lyophilized antiserum that reaches the certain purity Fab.
Polyvalent antiserum of the present invention has feature: with F (ab)
2Fab content is that 60%-80% lyophilizing porous solid shape antiserum makes up configuration, as table 1: table 1
| The snake venom kind | Agkistrodon halys | Agkistrodon | Adder | Ophiophagus hannan (Cantor) | Agkistrodon | Bungarus fasciatus | Naja |
| Antiserum consumption (mg/ person-portion) | 342.5-1035 | 1177-3182 | 456-666 | 1419-2128 | 220-331 | 880-1320 | 3188-4782 |
Antiserum optimum organization is configured to table 2: table 2
| The snake venom kind | Agkistrodon halys | Agkistrodon | Adder | Ophiophagus hannan (Cantor) | Agkistrodon | Bungarus fasciatus | Naja |
| Antiserum consumption (mg/ person-portion) | 635 | 1959 | 561 | 1774 | 276 | 1100 | 3985 |
The Ophiophagus hannan (Cantor) of the Agkistrodon halys of described blood circulation snake venom class, Agkistrodon, adder and neural snake venom class, Agkistrodon, Bungarus fasciatus, Naja antiserum be loading amount respectively.
Purified antivenin be by the blood plasma of immune animal through the digestion of pepsin short time, make its degeneration precipitation remove the Fc fragment that does not conform to antibody by heating, collection can with the bonded F of antigen (ab)
2Fragment and make F (ab)
2Fragment and antiserum combination will be determined in production and the standard in checking with magnitude relation.In the last table, contain 60%-80%F (ab) with antiserum
2Fragment is configured as the biomass concentration of nine kinds of snake venom of neutralization.After experiment showed, seven kinds of snake venom antiserum combination configurations, nine kinds of toxin expelling amounts that poisonous snake is common can neutralize.
Simultaneously, on immunology, same kind snake venom antibody-antigenic cross-reaction is stronger.Therefore, on the one hand, every loading amount of antivenin of the present invention is not only determined by the toxin expelling amount of purity height of antibody own and various poisonous snakes, is gone back fibrous root and calculate loading amount according to the amount of every kind of snake venom of serum cross-neutralization.On the other hand, neural malicious class and blood circulation poison class antiserum be loading amount respectively, can determine the Serpentis time-like, uses separately; When can not determine, answer two to use simultaneously, reduce and incur loss through delay.
Polyvalent antiserum using method of the present invention:
To the snake venom person that enters the body, with water for injection or normal saline dilution antivenin, muscle or intravenous injection are injected antiallergic agent simultaneously in the lump, as pounce on and quick or diphenhydramine or non-lagen or dexamethasone or strengthen can the ground pine.
The injection antiallergic agent is that this albumen is foreign protein to human body because the antivenin of animal serum preparation contains protein content more than 10%.When intravenous injection amount during up to 10ml, anaphylactoid incidence rate is eliminated fully.
The neural malicious class of loading amount and blood circulation poison class antiserum in the time determining the Serpentis kind, uses separately respectively; When can not determine, answer two to use simultaneously.
And the stability of antivenin and storage temperature are closely related.Even the lyophilizing antivenin is kept at 37 ℃, temperature is also not obvious to the influence of tiring, and is stored in below 4 ℃ to prolong sero-fast stability.
The present invention only uses the multivalence anti-venin serum of seven kinds of antivenins preparation, can be behind injection Claritin under the situation of not determining which kind of venom direct injection, treat in China region snakebite due to the various poisonous snakes in time, effectively.For this reason, make the antivenin quality of China reach international standard.
The specific embodiment
(1) loading amount of antivenin and stability
With content 60%-80% Fab F (ab)
2The sero-fast lyophilizing antiserum of Agkistrodon, Agkistrodon halys, adder, Agkistrodon, Bungarus fasciatus, Naja, Ophiophagus hannan (Cantor) make up this combination nine kinds of common toxin expelling amounts of poisonous snake that can neutralize.As table 3:
Table 3: the relation of antivenin loading amount and toxin expelling amount
By table 1 the lyophilizing antiserum of weighing is distributed into two of neural malicious class and blood circulation poison classes, is stored in below 4 ℃.
(2) polyvalent anti-snake poison lyophilized blood serum of the present invention is to nine kinds of snake venom effects of mice neutralization and dose-effect relationship
Tried mice and be divided into matched group and polyvalent serum administration group at random, every group more than 6.In the matched group, nine kinds of snake venom are pressed LD respectively
99Snake venom amount injection mouse peritoneal, the injection back was observed 72 hours.In the administration group, different antiserum dosage dissolve with the 0.2ml borate buffer solution, and nine kinds of LD again neutralize respectively
99The snake venom amount, behind 37 ℃ of water-bath 30min, the injection mouse peritoneal, 72 hours dead mouse situations are observed in the injection back.As table 4, with the polyvalent antiserum of various dose in nine kinds of snake venom and after, matched group mortality rate 100%, the mortality rate of maximum dose level group are 0%.
Table 4: multivalence anti-venin serum is in nine kinds of snake venom and mouse experiment and influence
(3) the clinical using method of antivenin of the present invention
Neural malicious class and blood circulation poison class antiserum to the difference loading amount in the time determining the Serpentis kind, use separately; When can not determine, answer two to use simultaneously.
In order to strive for treatment time, needn't make skin test when using antivenin.Elder generation intramuscular injection Claritin such as diphenhydramine 20mg also is diluted in 2 antivenins in the 200ml normal saline, and 15 of per minutes are done intravenous drip, after 15 minutes as reactionless, adds remaining all antivenins again and shakes up the back and do to instil fast.As the scalable drip velocity that responds is to reach desensitization effect.Counterweight danger patient should be than the big 3-6 of general dosage times or bigger.
Claims (3)
1. refined polyvalent anti-snake poison lyophilized blood serum is characterized in that with F (ab)
2Fab content is that 60%-80% lyophilizing porous solid shape antiserum makes up configuration, as following table:
2. refined polyvalent anti-snake poison lyophilized blood serum according to claim 1 is characterized in that antiserum optimum organization is configured to following table:
3. refined polyvalent anti-snake poison lyophilized blood serum according to claim 1 and 2 is characterized in that Agkistrodon halys, Agkistrodon, adder blood circulation poison class and Ophiophagus hannan (Cantor), Agkistrodon, Bungarus fasciatus, cobratoxin class antiserum loading amount respectively.
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| US11815311B2 (en) | 2019-03-14 | 2023-11-14 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101843902B (en) * | 2010-05-21 | 2012-03-07 | 成都军区疾病预防控制中心军事医学研究所 | Method for preparing polyvalent antiserum for resisting venin of common terrestrial viperidae venomous snakes in China and application thereof |
| CN106243224A (en) * | 2016-08-05 | 2016-12-21 | 安徽威尔试剂盒科技有限责任公司 | One utilizes Agkistrodon acutus venom differential protein to prepare sero-fast method and application thereof |
| CA3078625C (en) | 2017-10-09 | 2023-01-17 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
| CN110420325A (en) * | 2019-09-04 | 2019-11-08 | 沈龙平 | Refined polyvalent anti-snake poison lyophilized blood serum and preparation method thereof |
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| US4806346A (en) * | 1986-12-16 | 1989-02-21 | American Home Products Corporation | Method for isolation of antigen specific immunoglobulin |
| US6833131B1 (en) * | 1991-05-02 | 2004-12-21 | Protherics Inc. | Antivenom immunesera |
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| US4806346A (en) * | 1986-12-16 | 1989-02-21 | American Home Products Corporation | Method for isolation of antigen specific immunoglobulin |
| US6833131B1 (en) * | 1991-05-02 | 2004-12-21 | Protherics Inc. | Antivenom immunesera |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US11815311B2 (en) | 2019-03-14 | 2023-11-14 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
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Effective date of registration: 20160907 Address after: 315600, No. 39, Xincheng Road, Ningdong Town, Ninghai County, Zhejiang Province Patentee after: Zhejiang Jian Bo biological Polytron Technologies Inc Address before: 650032 No. 168 Daguan Road, Yunnan, Kunming Patentee before: Chengdu Military Command Sickness Prevention Control Centre War Medicine Institute |